Development of discovering technology to quantify the biomarker for minimally invasive diagnostics.

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The proteins they express at the time of disease and which are related to organ dysfunction are poorly characterized. Our group has developed a general chemical proteomics approach for the identification of biomarker proteins. The aim of this study was to identify protein biomarkers present in sinusoidal endothelial cells following hepatic injury using chemical proteomics employing in vivo biotinylation. We performed a proteomic study in a mouse model of massive hepatocyte apoptosis in tumor necrosis factor (TNF)/D-(+)-galactosamine (GalN) animals induced by the combined administration of GalN and TNF. In vivo biotinylation was carried out by perfusing mice with a reactive ester derivative of biotin that enables the covalent modification of proteins. Protein expression was analyzed using purified proteins from hepatitic and normal livers. The biotinylated proteins were purified from liver extracts using streptavidin beads, digested on the resin by trypsin and subjected to mass spectrometry for identification. These results indicate that the sinusoidal endothelial biomarkers expressed on liver injury reflect the molecular pathobiology and the mechanisms of hepatitis induction.